Validation of a method for determination of hemagglutinin content in quadrivalent influenza vaccine (split virion) inactivated

Objective To validate the feasibility of a method for determination of hemagglutinin content in quadrivalent influenza vaccine(split virion)inactivated(QIV). Methods The contents of hemagglutinin of subtypes H1N1 and H3N2 in QIV were determined by routine single-radial immunodiffusion(SRID)test, while those of subtype B(B / Yam and B /Vic) by bivalent antigen reference SRID test. The method was validated for accuracy, precision, specificity and linearity.Results The total recovery rates of hemagglutinin contents of various subtypes were 97. 8% ~ 107. 6%, with relative standard deviations(RSDs)of 0. 59% ~ 1. 83%. Both RSDs in inter- and intra-assays were less than 5. 0%. The standard curve showed good linearity at hemagglutinin contents of four subtypes of 10 ~ 50 μg/ml, with R2 values of more than0. 980 0. The quantitative limits of subtypes H1N1, H3N2, B / Yam and B / Vic were 4. 53, 3. 83, 2. 89 and 5.60 μg/ml respectively. Both vaccine samples and antigen references of various subtypes showed precipitation rings with the corresponding antiserum references, while showed no precipitation rings with non-corresponding antiserum references.Conclusion The method for determination of hemagglutinin content in QIV was feasible.